Since the 1994 DSHEA mandate, botanical dietary supplements (BDSs) are continuously developing into safer and more effective preparations that aid U.S. public health. Since its establishment in 1999, our BOTANICAL CENTER has participated in this development by performing significant (bio-)chemical and clinical research on BDSs that are widely used to restore and/or maintain women's health. The focus of a renewed grant period will continue to be the alleviation of perimenopausal and premenstrual syndrome (PMS) symptoms. Linked to this focus is the central hypothesis that botanicals contain potent secondary plant metabolites that exhibit activities in estrogenic and major CNS (serotonin, GAB A, dopamine, and opioid) systems. In close collaboration with the other projects of the CENTER, PROJECT by Farnsworth will (Aim 1) evaluate and prioritize select botanicals; (Aim 2) show that they contain phytoconstituents with the desired biological activities and profiles, in vitro and in vivo; isolate and structurally characterize these active principles by modern spectroscopical methods and use them to (Aim 3) standardize the botanicals. The long-term objective of this proposal is to prepare botanical extracts that are chemically and biologically standardized in terms of the bioactive secondary plant metabolite(s), and that are stable for human studies. Combining promising botanicals from the previous grant period with the results of preliminary studies, an initial selection of 12 plants plus two complex herbal mixtures will be evaluated. Primary extracts of a wide polarity range and their solvent partition fractions will be assayed in vitro and in vivo by PROJECTS by Bolton and prioritized using a scoring system that weighs hit numbers, grouping of potency, as well as strengths and concentration of activity. The botanicals with the highest rank will be scaled-up and subjected to an innovative bioassay-guided fractionation procedure (w/PROJECTS by Bolton and Analytical Core). The major active principles will be obtained through a sequential application of modern chromatography tailored to the specific characteristics of the extracts (HSCCC, Gel/Resin-CC). Modern qualitative and quantitative spectroscopic analysis (1/2D and selective NMR, qNMR, hyphenated MS) will lead to the identification of structures of the major active principles, which ultimately lays the foundation for chemical and biological standardization. Prioritized botanicals will be fully developed into standardized products. Hops will be evaluated in a Phase I clinical study in PROJECT 4.